The invasion assay was performed as reported previously with some modifications [45 (link)]. The cells' invasive and migration abilities were examined using a 24-well trans-well with 8-μm pore polycarbonate membrane inserts (Becton Dickinson, Franklin Lakes, NJ, USA) according to the manufacturer's protocol. 2× 106 cells were planted on the upper chamber. After 48h in culture, cells that appeared on the lower surface of the filter were fixed with 100% methanol and stained with crystal violet and counted in five random ×200 fields using an inverted microscope and with the double-blind method. The experiments were performed in triplicate and got means were calculated.
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